title: Cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of Rv0241c (HtdX) from Mycobacterium tuberculosis H37Rv.
abstract: Rv0241c (HtdX) is a putative (3R)-hydroxyacyl-CoA dehydratase of Mycobacterium tuberculosis. The htdX gene belongs to a conserved operon and is expressed in mycobacteria in the presence of several fatty-acid synthase II drugs. To elucidate the structure of HtdX, the protein was cloned, overexpressed, purified to homogeneity and crystallized. The protein was crystallized from two conditions: (i) 3 M sodium chloride, 0.1 M Na HEPES pH 8.0 and (ii) 2.5 M sodium chloride, 0.1 M Tris-HCl pH 8.5. A complete diffraction data set was collected from crystals from both conditions. The crystal from the first condition diffracted to 2.3 Å resolution and belonged to space group I41, with unit-cell parameters a=b=61.51, c=143.81 Å. Crystals from the second condition diffracted to 3.1 Å resolution and belonged to space group P4₃2₁2 or P4₁2₁2, with unit-cell parameters a=b=63.67, c=140.88 Å. Both crystals contained one molecule in the asymmetric unit.
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721 302, India.
authors.forename: Rupam
authors.initials: R
authors.lastname: Biswas
authors.forename: Debajyoti
authors.initials: D
authors.lastname: Dutta
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Das
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D003001:Cloning, Molecular; D003460:Crystallization; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins
keywords: Mycobacterium tuberculosis; Rv0241c (HtdX)